Early Pregnancy Period Research Articles

Relationship between hypertensive disease stages of pregnancy and plateletcrit values: A retrospective cohort study

ObjectiveThe aim of this study was to investigate the correlation between plateletcrit (PCT) and stages of hypertensive disease of pregnancy by comparing values in groups with healthy pregnancies, gestational hypertension [GHT], preeclampsia with proteinuria [PE + P] and severe preeclampsia with proteinuria [sPE + P]. The secondary aim was to investigate how proteinuria affects PCT values in these patients. Materials and methodsThis was a retrospective cohort study. Patients who had been hospitalized for labor were included in the study. The following groups were created: healthy pregnancies (n = 36), [GHT] (n = 36), [PE + P] (n = 36) and [sPE + P] (n = 36). PCT and platelet values from blood tests performed immediately after hospitalization were compared among the four groups. Additionally, the PCT and platelet values in the early pregnancy period of same patients were determined and compared. ResultsThere were no statistically significant differences among the groups in terms of age, gravidity or parity (p > 0.05). The mean platelet counts of the groups ranged from 219,250/mm3 to 241,310/mm3. There were no significant differences among the groups (with or without proteinuria) in terms of the platelet counts or PCT values (p > 0.05). A reducing or enhancing effect of proteinuria on the PCT value was not observed. Additionally, based on data from the early weeks of pregnancy, there were no significant differences among the groups in terms of the platelet counts or PCT values (p > 0.05). ConclusionsPCT may not be appropriate to use for the diagnosis or prediction of sPE in patients whose platelet counts have not decreased. A normal PCT value should not be interpreted as the absence of progression to sPE. There is a need for new PCT and PE studies that take platelet counts into account and include women in the early weeks of pregnancy.

Preconception and Early-Pregnancy Body Mass Index in Women and Men, Time to Pregnancy, and Risk of Miscarriage

Obesity in women is associated with reduced fertility and an increased risk of miscarriage. These associations might also be present across the full range of body mass index (BMI) categories as well as among men. To investigate the associations of preconception BMI in both partners with time to pregnancy and miscarriage. This population-based prospective cohort study was conducted in Rotterdam, the Netherlands, between August 9, 2017, and July 1, 2021. A total of 3604 women and their partners were included from the preconception period onward with follow-up until birth. The date of analysis was July 12, 2024. Body mass index (calculated as weight in kilograms divided by height in meters squared) measured in preconception or early pregnancy. Fecundability, defined as the probability of conceiving within 1 month; subfertility, defined as time to pregnancy or duration of actively pursuing pregnancy of more than 12 months or use of assisted reproductive technology; and miscarriage, defined as pregnancy loss before 22 weeks of gestation. These measures were assessed using questionnaires and via the obstetric caregiver. The study population for time-to-pregnancy analyses consisted of 3033 episodes among women (median age, 31.6 years [IQR, 29.2-34.5 years]; median BMI, 23.5 [IQR, 21.2-26.5]) and 2288 episodes among men (median age, 33.4 years [IQR, 30.5-36.8 years]; median BMI, 24.9 [IQR, 23.0-27.4]). The study population for miscarriage analyses consisted of 2770 pregnancy episodes among women (median age, 31.5 years [IQR, 28.9-34.3 years]; median BMI, 23.5 [IQR, 21.3-26.7]) and 2189 pregnancy episodes among men (median age, 33.5 years [IQR, 30.4-36.8 years]; median BMI, 25.0 [IQR, 23.0-27.5]). Higher BMI in women and men was associated with lower fecundability: for every unit increase in BMI, fecundability decreased (fecundability ratio [FR]: women, 0.98 [95% CI, 0.97-0.99]; men, 0.99 [95% CI, 0.98-1.00]). Women with overweight (FR, 0.88 [95% CI, 0.80-0.98]) and obesity (FR, 0.72 [95% CI, 0.63-0.82]) had lower fecundability compared with women with normal weight. Compared with normal weight in women, underweight (odds ratio [OR], 1.88 [95% CI, 1.22-2.88]), overweight (OR, 1.35 [95% CI, 1.11-1.63]), and obesity (OR, 1.67 [95% CI, 1.30-2.13]) were associated with increased odds of subfertility. In men, obesity was associated with increased odds of subfertility (OR, 1.69 [95% CI, 1.24-2.31]). Compared with normal weight in women, overweight (OR, 1.49 [95% CI, 1.12-1.98]) and obesity (OR, 1.44 [95% CI, 1.00-2.08]) were associated with increased odds of miscarriage. In this cohort study, BMI outside of the normal category in women and men during the preconception or early-pregnancy periods was associated with time to pregnancy and miscarriage. Optimizing BMI in women and men from the preconception period onward might be an important strategy to improve fertility outcomes.

Climate Change and Congenital Anomalies: A Population-Based Study of the Effect of Prolonged Extreme Heat Exposure on the Risk of Neural Tube Defects in France.

Exposure to long-lasting extreme ambient temperatures in the periconceptional or early pregnancy period might increase the risk of neural tube defects (NTDs). We tested whether prolonged severe heat exposure as experienced during the 2003 extreme heatwave in France, affected the risk of NTDs. We retrieved NTD cases spanning from January 1994 to December 2018 from the Paris Registry of Congenital Malformations. The 2003 heatwave was characterized by the long duration and high intensity of nine consecutive days with temperatures ≥35°C. We classified monthly conceptions occurring in August 2003 as "exposed" to prolonged extreme heat around conception (i.e., periconceptional period). We assessed whether the risk of NTDs among cohorts exposed to the prolonged severe heatwave of 2003 in the periconceptional period differed from expected values using Poisson/negative binomial regression. We identified 1272 NTD cases from January 1994 to December 2018, yielding a monthly mean count of 4.24. Ten NTD cases occurred among births conceived in August 2003. The risk of NTD was increased in the cohort with periconceptional exposure to the August 2003 heatwave (relative risk = 2.14, 95% confidence interval: 1.46 to 3.13), compared to non-exposed cohorts. Sensitivity analyses excluding July and September months or restricting to summer months yielded consistent findings. Evidence from the "natural experiment" of an extreme climate event suggests an elevated risk of NTDs following exposure to prolonged extreme heat during the periconceptional period.

Cardiac remodelling during pregnancy in women with congenital heart disease and systemic left ventricle.

Women with congenital heart disease (CHD) are at risk of pregnancy-related adverse outcomes (PRAO). The purpose of this study was to assess temporal changes in cardiac structure and function (cardiac remodelling) during pregnancy, and the association with PRAO in women with CHD. Retrospective study of pregnant women with CHD and serial echocardiograms (2003-2021). Cardiac structure and function were assessed at pre-specified time points: prepregnancy, early pregnancy, late pregnancy, and postnatal period. PRAO was defined as the composite of maternal cardiovascular, obstetric, and neonatal complications. The study comprised 81 women with CHD (age, 29 ± 5 years). Compared to the baseline echocardiogram, there was a relative increase in right ventricular systolic pressure (RVSP) (relative change 13 ± 5%, P < 0.001, in early pregnancy; and 18 ± 5%, P < 0.001, in late pregnancy). There was a relative decrease in right ventricle free wall strain (RVFWS) (relative change -11 ± 3%, P < 0.001, in late pregnancy; and -11 ± 4%, P = 0.003, in postnatal period), and a relative decrease in RVFWS/RVSP (relative change, -10 ± 5%, P = 0.02 in early pregnancy, -26 ± 7%, P < 0.001, in late pregnancy, and -14 ± 5%, P < 0.001, in postnatal period). Baseline right ventricular to pulmonary arterial (RV-PA) coupling, and temporal change in RV-PA coupling were associated with PRAO, after adjustment for maternal age and severity of cardiovascular disease. Women with CHD had a temporal decrease in RV systolic function and RV-PA coupling, and these changes were associated with PRAO. Further studies are required to delineate the aetiology of deterioration in RV-PA coupling during pregnancy, and the long-term implications of right heart dysfunction observed in the postnatal period.

Determinants of neural tube defect among newborns admitted to neonatal intensive care units of teaching hospitals in Gedeo Zone and Sidama Region, Southern Ethiopia: a case–control study

BackgroundNeural tube defects are a significant cause of morbidity and mortality that can occur in the early pregnancy periods. Though the burden is high, it gains only limited attention. In...

Associations of COVID-19 lockdown with birth weight in China.

During the special period of the global spread of COVID-19, pregnant women are sensitive groups to the impacts of COVID-19 epidemic. However, the effects of lockdown measures implemented in response to the COVID-19 on fetal birthweight remain unclear. This study investigated the associations of COVID-19 lockdown with birth weight in Chinese population. We collected 730,153 data of participants from hospitals of five cities in the south of China, we defined the time period of level I response (1/23-2/24/2020) as level I lockdown, and women who were pregnant during level I lockdown as the exposure group. Women who were pregnant during the same calendar month from 2015 to 2019 were defined as the unexposed group. We quantitatively estimate the individual cumulative exposure dose by giving different weights to days with different emergency response levels. Generalized linear regression models were used to estimate the association between COVID-19 lockdown exposure with birth weight and risk of low birth weight (<2,500 g) and macrosomia (>4,000 g). The birth weight of the exposed group is heavier than the unexposed group (3,238.52 vs. 3,224.11 g: adjusted β = 24.39 g [95% CI: 21.88, 26.91 g]). The exposed group had a higher risk of macrosomia (2.8% vs. 2.6%; adjusted OR = 1.17 [95% CI: 1.12, 1.22]). More obvious associations were found between COVID-19 lockdown and macrosomia in women who experienced the lockdown in their early pregnancy. Women who experienced the lockdown at their 4-7 weeks of pregnancy showed statistically significant heavier birth weight than unexposed group (after adjustment): β = 1.28 (95% CI: 1.11, 1.46) g. We also observed a positive association between cumulative exposure dose of COVID-19 lockdown in all pregnant women and birth weight, after divided into four groups, Q1: β = 32.95 (95% CI: 28.16, 37.75) g; Q2: β = 18.88 (95% CI: 14.12, 23.64) g; Q3: β = 19.50 (95% CI: 14.73, 24.28) g; Q4: β = 21.82 (95% CI: 17.08, 26.56) g. However, there was no statistically significant difference in the risk of low birth weight between exposed and unexposed groups. The COVID-19 lockdown measures were associated with a heavier birth weight and a higher risk of macrosomia. Early pregnancy periods may be a more susceptible exposure window for a heavier birth weight and a higher risk of macrosomia. We also observed a positive association between cumulative exposure dose of COVID-19 lockdown and birth weight. The government and health institutions should pay attention to the long-term health of the infants born during the COVID-19 lockdown period, and follow up these mothers and infants is necessary.

Preconceptional micronutrient adequacy among women in Greece: a prospective epidemiological study

Introduction The importance of micronutrient intake during the preconceptional and early pregnancy period for both maternal and fetal outcomes is well-known, however, relevant data are not available for Greek pregnant women. The aim of the present study is to delineate the nutritional status preceding conception among a representative cohort of Greek pregnant women. Methods This was a prospective study of pregnant women from routine care, recruited at 11+0–13+6 gestational weeks, between December 2020 and October 2022, at the 3rd Department of Obstetrics and Gynecology, Aristotle University of Thessaloniki, Greece. Eligible participants for the study included healthy pregnant women aged 20 years or older, possessing a proficient understanding of the Greek language, and not engaged in specific nutritional programs. A validated Food Frequency Questionnaire was applied to gather information regarding nutritional habits in the last 6 months prior to conception. The consumption of nutrients was compared to the reference intake levels suggested by the European Food Safety Authority. Further analyses between different participants’ subgroups were performed. Results Overall, 1100 pregnant women (mean age: 32.4 ± 4.9 years) were enrolled. Almost all examined micronutrients’ intake was significantly different from dietary reference values. Furthermore, nutrient adequacy ratio was below 60% in 6 out of 22 micronutrients examined, and Mean Adequacy Ratio was 93%. However, Mean Adequacy Ratio is characterized by extreme variance between the examined values. Iodine, folic acid, potassium, and vitamin D intake levels were significantly lower than the recommended intake levels (p < .001 for all), while vitamin K and niacin (p < .001 for both) were consumed in great extent. Sodium median intake, without calculating extra salt addition also exceeded the reference value levels (p = .03). Notably, magnesium intake exceeded the upper safety limits in 12.4% of the sample. Conclusion Potential inadequacies in important micronutrients for uneventful pregnancy outcomes have been revealed.. Special attention is needed for magnesium to balance possible toxicity with evident benefits.

Association of Maternal Serum Gamma-Glutamyltransfetase Level at First Trimester as a Predictive Biomaker of Gestational Diabetes Mellitus

Introduction: The prevalence of GDM is increasing globally. GDM is associated with many maternal and fetal complications. Hence identifying and treating maternal hyperglycemia is beneficial for both mother and baby. Proper monitoring of women is necessary during pre-conceptional and early pregnancy period to prevent GDM and its complication. Objective: To assess the level of maternal serum Gamma-glutamyltransferase (GGT) as predictor of Gestational Diabetes Mellitus (GDM). Methods: A Prospective cohort study was carried out at Department of Obstetrics and Gynaecology, Dhaka Medical College Hospital, Dhaka from January 2021 to December 2021. Total sample size was 144. Pregnant women attending Obstetric OPD, DMCH at her 1st trimester (8 -12 weeks) without the evidence of GDM, DM, or any comorbid condition and drugs that affects blood glucose level. Data regarding demographic profile, medical and family history (history of chronic hypertension, diabetes mellitus, autoimmune disease, and CKD) were recorded. Obstetric history regarding gravity, parity, past obstetric history was documented. Then a complete physical examination of the subjects including height, weight, blood pressure level was carried out with an appropriate procedure. Period of the gestation was confirmed by recording CRL in early USG. Patient’s fasting blood sugar and 2hrs after 75- gram glucose at first trimester was measured. Results: In the current study 144 pregnant women, at their first trimester (8-12 week) were included, but during follow up about 12 patients were dropped out due to mid trimester abortion, migration to another place and so on. Most of them were primi (56.1%) housewife (75%), had completed their secondary education (59.1%), and about 61.4% belong to middle class family. The clinical parameter like BMI 25.80±4.56kg/m2, gestational age at delivery 38.09±1.28 weeks. The mean ±SD BMI 27.52±4.72 kg/m in GDM group 25.40±4.46 kg/m in Non GDM group. The mean ± SD birth weight was 3.07±0.37 kg in GDM group and 2.86±0.33 in Non GDM group. The difference was statistically non- significant (p&gt;0.05) between two groups. Among 45.45% subjects had GGT level &lt;10 U/L,34.09% had 11-20 U/L and 20.45% had GGT &gt;20 U/L. Among total 132 respondents 25 respondents (18.94%) developed GDM. The total population divided in two groups, Group I (GDM) and Group II (non GDM). Shows that in group A (GGT ≥12.5 U/L) total respondents were 61, GDM developed in 29.50% (n=18), GDM not developed in 70.49% (n=43). On the Other hand in Group B (GGT &lt;12.5 U/L) total respondents were 71, GDM developed in 9.85% (n=07) and GDM not developed in 90.14% (n=64) and p value was 0.004. Conclusion: The result of our study indicated that pregnant women with high GGT (&gt;12.5 U/L) at booking visit had greater risk of developing GDM (72%).Sensitivity, Specificity, PPV ,NPV of this diagnostic test was 72% , 59.81%, 29.50%, 90.14% respectively and accuracy is 94.69%. Among these 132 respondents, those having GGT&gt;12.5IU/L relative risk (RR) of development of GDM is 3.01. As measurement of GGT is easy, inexpensive and easy to carry out, it can be used as a predictive biomarker who are at increased risk for the subsequent development of GDM.

The Effect of Human Chorionic Gonadotropin and Ketoprofen Applications on Pregnancy Rates in Dairy Cows After Artificial Insemination

The aim of the presented study is to investigate the effect of providing progesterone (P4) support for the formation and maintenance of pregnancy or suppressing the luteolysis mechanism during the maternal acceptance process on pregnancy rates. For this purpose, 96 Holstein cows, aged between 2 and 8 years, were synchronized using the Ovsynch protocol to align their estrous cycles. After artificial insemination, the cows were divided into four groups. The group receiving Human Chorionic Gonadotropn (hCG) on the 6th day after artificial insemination (H; n=20), the group receiving Ketoprofen on the evening of the 15th day and the morning of the 16th day after artificial insemination (K; n=24), the group receiving both hCG on the 6th day and Ketoprofen on the 15th day evening and 16th day morning after artificial insemination (HK; n=25), and the control group (C; n=27) that did not receive any post-insemination treatment were established. Pregnancy checks were conducted on the 30th, 45th, and 60th days through rectal and ultrasonographic (USG) examinations. According to the results of the pregnancy examinations, it was determined that 9 out of 20 cows (45.00%) in Group H, 10 out of 24 cows (41.67%) in Group K, 16 out of 25 cows (64.00%) in Group HK, and 12 out of 27 cows (44.44%) in Group C were pregnant. No statistically significant difference (P&gt;0.05) was observed among the groups. Our results suggest that a multi-purpose treatment may be required to reduce pregnancy loss during the early pregnancy period in cows due to an approximately 20% increased pregnancy rate after administering hCG and Ketoprofen. However, further studies, especially involving more cows, are necessary to optimize and use these combined treatments and potentially increase pregnancy rates.

Different seminal ejaculated fractions in artificial insemination condition the protein cargo of oviductal and uterine extracellular vesicles in pig.

The seminal plasma (SP) is the liquid component of semen that facilitates sperm transport through the female genital tract. SP modulates the activity of the ovary, oviductal environment and uterine function during the periovulatory and early pregnancy period. Extracellular vesicles (EVs) secreted in the oviduct (oEVs) and uterus (uEVs) have been shown to influence the expression of endometrial genes that regulate fertilization and early embryo development. In some species, semen is composed of well-separated fractions that vary in concentration of spermatozoa and SP composition and volume. This study aimed to investigate the impact of different accumulative fractions of the porcine ejaculate (F1, composed of the sperm-rich fraction, SRF; F2, composed of F1 plus the intermediate fraction; F3, composed of F2 plus the post-SRF) on oEVs and uEVs protein cargo. Six days after the onset of estrus, we determined the oEVs and uEVs size and protein concentration in pregnant sows by artificial insemination (AI-sows) and in non-inseminated sows as control (C-sows). We also identified the main proteins in oEVs and uEVs, in AI-F1, AI-F2, AI-F3, and C-sows. Our results indicated that although the size of EVs is similar between AI- and C-sows, the protein concentration of both oEVs and uEVs was significantly lower in AI-sows (p < 0.05). Proteomic analysis identified 38 unique proteins in oEVs from AI-sows, mainly involved in protein stabilization, glycolytic and carbohydrate processes. The uEVs from AI-sows showed the presence of 43 unique proteins, including already-known fertility-related proteins (EZR, HSPAA901, PDS). We also demonstrated that the protein composition of oEVs and uEVs differed depending on the seminal fraction(s) inseminated (F1, F2, or F3). In conclusion, we found specific protein cargo in oEVs and uEVs according to the type of semen fraction the sow was inseminated with and whose functions these specific EVs proteins are closely associated with reproductive processes.

High initial β-hCG predicts IVF outcomes accurately and precludes the need for repeated measurements.

This study evaluated β-hCG changes during the early period of pregnancy in an attempt to predict successful pregnancy outcomes in ART. It determined the median values of the β-hCG and the 2-day β-hCG increments of clinical vs. biochemical pregnancies. The results of fresh day 3 embryo, frozen day 3 embryo and frozen day 5 embryo transfers were evaluated. The cutoff values of β-hCG and the 2-day increments predicting clinical pregnancy and delivery were determined. All women who underwent embryo transfer and had a singleton pregnancy from January 2017-December 2019 were included. As expected, clinical pregnancies had higher initial median β-hCG values compared to biochemical pregnancies (fresh day 3 (400 vs. 73 mIU/ml), frozen day 3 (600 vs. 268.5 mIU/ml) and frozen day 5 (937 vs. 317 mIU/ml). Nonetheless, the abortion rate was significantly lower in the group with β-hCG above the cutoff values in fresh (141 mIU/ml), and frozen (354.5 mIU/ml) cleavage stage transfers (17.2% vs. 44%, P<0.001 and 18.5% vs. 38%, P=0.003, respectively). Blastocyst transfers resulted in higher median initial β-hCG compared to cleavage embryo transfers (937 vs. 600 mIU/ml), and the initial β-hCG values from frozen cleavage embryos were higher compared to fresh cleavage embryos (600 vs. 400 mIU/ml). Earlier implantation in frozen cycles may be caused by freezing-thawing procedures. Moreover, in fresh cycles, negative effects of the hormonal milieu of fresh cycles may delay implantation. Results indicate that high initial β-hCG and high 2-day β-hCG increments demonstrated better outcomes, including more clinical pregnancies and fewer abortions.

Th1 or Th2 cytokines are correlated with Tregs and T cell subsets and pregnancy outcomes in patients with autoimmune thyroid disease during early, middle, late pregnancy, and postpartum period

Autoimmune thyroid disease (AITD) is a T lymphocytes-mediated autoimmune disorder affecting pregnant women. The current study sought to determine the correlations between T helper-1 (Th1)/T helper-2 (Th2) cytokines and regulatory T cells (Tregs) and T cell subsets and pregnancy outcomes in AITD patients during early pregnancy (T1), middle pregnancy (T2), late pregnancy (T3), and postpartum period (PP). A total of 60 patients with Graves’ disease, 60 patients with Hashimoto’s thyroiditis, and 30 healthy pregnant women were initially enrolled in the study. Thyroid hormones and antibodies, Th1 or Th2 cytokines, transforming growth factor-β, Tregs, CD4+ T helper cells (CD4+), CD8+ T helper cells (CD8+) levels were determined by means of Maglumi2000 automatic chemiluminescence instrument, enzyme-linked immunosorbent assay, and flow cytometry. Our findings demonstrated higher IFN-γ and IL-2 levels, along with lower IL-4, IL-10, TGF-β, Treg, and CD4+/CD8+ levels in AITD patients during T1, T2, T3, and PP. Furthermore, the TGF-β, Treg, and CD4+/CD8+ levels were lower in the IFN-γ/IL-2 high expression group but higher in the IL-4/IL-10 high expression group. The IFN-γ and IL-2 levels were higher, while IL-4 and IL-10 level were lower in AITD patients with adverse pregnancy outcomes. Lastly, Th1 cytokines were higher and Th2 cytokines were lower in AITD patients and elicited correlation with Tregs and CD4+/CD8+ levels. Collectively, our findings highlighted that up-regulation of Th1 cytokines may increase the percentage of adverse pregnancy outcomes in AITD patients.

Impact of ambient temperature on adverse pregnancy outcomes: a birth cohort study in Fuzhou, China.

Previous studies have identified a series of specific adverse pregnancy outcomes (APOs) linked with temperature extremes. Most of them focus on preterm birth, low birth weight, and stillbirth. Other possible adverse outcomes were under-researched. This study aimed to investigate the impact of ambient temperature on maternal complications, white blood cell count (WBC), newborn hearing, and neonatal jaundice. A total of 418 participants were recruited from Fuzhou Maternity & Child Healthcare Hospital in 2016. Participants were invited to fill out a structured questionnaire. The gridded near-surface air temperatures at a resolution of 0.1°* 0.1° for Fuzhou were extracted from a published dataset. Meteorological data and PM2.5 were extracted based on participants' residential addresses using R packages "ncdf4" and "raster." Multivariate logistic regression models were used to quantify the effects of ambient temperature on APOs after controlling for confounders. Overall, there were 107 APOs, accounting for 25.6% of all participants. Every 1°C increase in mean temperature was associated with a 10.0% increase in APOs (aOR = 1.100, 95%CI 1.006-1.203) during the period of early pregnancy. However, negative associations were observed in the middle pregnancy period, and a 1°C increase in mean temperature was associated 8.8% decrease in APOs (aOR = 0.912, 95%CI 0.846-0.982). Diurnal temperature variation had a significant impact on APOs in the third trimester. Infant jaundice was negatively associated with temperature exposure in the middle and late pregnancy periods. The risk of neonatal jaundice increased at lag weeks 2-9 in the first trimester, with the greatest lagged effect (aOR = 1.201, 95%CI 1.020-1.413) observed at lag week 3. A 1°C increase in mean temperature led to a 29.6% (aOR = 1.296, 95%CI 1.019-1.649) increase in high WBC. A 1°C increase in temperature variation was associated with more than two times (aOR = 2.469, 95%CI 1.001-6.089) increase of high WBC in the first trimester and about five times (aOR = 4.724, 95%CI 1.548-14.409) increase in the third trimester. Ambient temperature affects neonatal jaundice, newborn hearing loss, and infections during pregnancy. In addition to the identified epidemiologic link and susceptible exposure windows, there is a need to understand the underlying biological mechanisms for better recommendations for climate change adaptation policies.

Embryo-maternal interactions revealed by phosphoproteomic analysis of endometrial organoids exposed to embryo secretome

The equine embryo remains mobile and without physical attachment to the endometrium for a prolonged period in early pregnancy. During this time, the embryo only communicates with the maternal system via secreted compounds in its immediate environment. These interactions are poorly understood, and a definitive signal for maternal recognition of pregnancy remains elusive. In this study, we sought to develop a model to examine embryo-maternal interactions at close range in vitro, demonstrating that incubation of endometrial organoids with embryo-conditioned medium results in detectable changes in the endometrial phosphoproteome. Mare endometrial organoids were derived from isolated endometrial glands of uteri obtained immediately post-mortem. Embryos were recovered by uterine lavage from Standardbred mares at day 8-9 post-ovulation, washed and pre-cultured in BSA-containing medium under oil, and cultured in protein-free medium under oil. Embryo-conditioned and control media were collected at 12 h, centrifuged to remove debris and stored at −80°C until further analysis. For co-incubation experiments, organoids were primed with 10nM estradiol (48 h), then 10nM estradiol + 1 µM progesterone (48 h). Embryo-conditioned (n=3) or control (n=3) media were added at a dilution of 1:100 in organoid medium and co-incubated for 30 minutes. Organoids were then recovered and processed to remove media and secretome components (on ice and in the presence of phosphatase inhibitors). Cell pellets were frozen at -80˚C until protein extraction. Proteins were extracted with urea-based extraction buffer, reduced,alkylated, and digested with trypsin. Phosphopeptides were isolated using titanium dioxide before LC tandem mass spectrometry (MS/MS) using a Q-Exactive Plus hybrid quadrupole-Orbitrap MS system (Thermo Fisher Scientific, Bremen, Germany). Endometrial organoids were successfully cultured in vitro and their functional mimicry of in vivo endometrium was confirmed via multiple routes including histological analysis of gland-like structures, epithelial and endometrial markers, and RNA sequencing alongside isolated endometrial glands. Phosphoproteomic mass spectrometry of endometrial organoids revealed a profile of 181 phosphopeptides mapping to 90 protein IDs. Of these, 10 phosphopeptides were decreased (log2[fold-change] < -0.8) in co-incubated organoids, while 3 were uniquely detected in secretome-exposed organoids only. Most notably, we observed a decrease in the phosphorylated peptides of PGRMC1 (progesterone receptor membrane component 1). This protein forms part of a non-classical system of progesterone action, located in the cell membrane. A change in PGRMC1 phosphorylation suggests that an alternative system of progesterone receptor activation participates in equine embryo-maternal signaling. There were also changes in phosphorylation status of chaperone proteins essential to progesterone receptor assembly (e.g., HSP90AB1). In summary, this study describes the immediate response of mare endometrial organoids to compounds released by embryos, identifying novel mechanisms possibly implicated in signaling between the embryo and maternal system.

Exposure to ambient air pollution from the preconceptional period and risk of gestational hypertension

Although accumulative studies have revealed the associations between air pollutants and elevated risk of gestational hypertension (GH), evidence from developing countries with relatively higher levels of air pollutants remains limited. In this retrospective study, a total of 45,439 birth records were collected in Beijing, China from 2013 to 2018. For PM2.5, SO2, NO2, and O3, exposure windows from the 3rd month of preconception to the 6th month of conception and the averages of 3 months of preconception, trimester 1 and trimester 2 periods were all calculated for assessment of GH risks. The correlations between air pollutants and the risk of GH were analyzed by logistic regression model. Our results showed that exposure to PM2.5 and SO2 in the preconceptional and early pregnancy periods was related to the elevated risk of GH. Furthermore, 3 months preconceptional exposure to PM2.5 (PCPM2.5: OR = 1.134 (1.114, 1.155)) and SO2 (PCSO2: OR = 1.158 (1.135, 1.181)) showed a higher risk of GH than the results of the trimester 1 (T1PM2.5: OR = 1.131 (1.104, 1.159); T1SO2: OR = 1.164 (1.141, 1.187)) and the trimester 2 (T2PM2.5: OR = 1.154 (1.126, 1.182); T2SO2: OR = 1.121 (1.098, 1.144)). The study also found significant and higher OR values for PCPM2.5, and PCSO2 from 2013 to 2016 when air pollution was serious in Beijing compared with 2017 to 2018 when the air pollution was obviously improved. Subgroup analysis also found that during 3 months of preconception women with higher age and who exposure to higher temperatures showed higher GH risk from PM2.5 and SO2 than that of the younger group and who exposure to lower temperature, respectively. Collectively, our findings suggest that air pollution exposure was adversely associated with GH in pregnant women and the preconceptional period is a critical air pollution exposure window for GH. Improving air quality can benefit public health, especially for sensitive populations like pregnant women.

Association between frequency of breakfast intake before and during pregnancy and infant birth weight: the Tohoku Medical Megabank Project Birth and Three-Generation Cohort Study

BackgroundLow birth weight is associated with an increased risk of developing chronic diseases in adulthood, with a particularly high incidence in Japan among developed countries. Maternal undernutrition is a risk factor for low birth weight, but the association between the timing of food intake and infant birth weight has not been investigated. This study aimed to examine the association between breakfast intake frequency among Japanese pregnant women and infant birth weight.MethodsOf all pregnant women who participated in the Tohoku Medical Megabank Project Three Generation Cohort Study, 16,820 who answered the required questions were included in the analysis. The frequency of breakfast intake from pre- to early pregnancy and from early to mid-pregnancy was classified into four groups: every day and 5–6, 3–4, and 0–2 times/week. Multivariate linear regression models were constructed to examine the association between breakfast intake frequency among pregnant women and infant birth weight.ResultsThe percentage of pregnant women who consumed breakfast daily was 74% in the pre- to early pregnancy period and 79% in the early to mid-pregnancy period. The average infant birth weight was 3,071 g. Compared to women who had breakfast daily from pre- to early pregnancy, those who had breakfast 0–2 times/week had lower infant birth weight (β = -38.2, 95% confidence interval [CI]: -56.5, -20.0). Similarly, compared to women who had breakfast daily from early to mid-pregnancy, those who had breakfast 0–2 times/week had lower infant birth weight (β = -41.5, 95% CI: -63.3, -19.6).ConclusionsLess frequent breakfast intake before and mid-pregnancy was associated with lower infant birth weight.

First Evidence on the Adverse Effect of Maternal Germanium Exposure on Fetal Neural Tube Defects

Germanium (Ge) can induce certain neurotoxic effects and may affect the fetal neural tube development. We aimed to investigate its association with the risk of neural tube defects (NTDs) by adopting integrated evidence. The potential associations between maternal hair Ge concentrations were explored using a relatively large case–control study of 285 NTD cases (women delivering a birth affected by NTDs) and 543 controls (those delivering healthy fetus without NTDs). The potential toxicological pathways and the reliability of the exposure biomarker of hair Ge were confirmed using additional population study and rat model, respectively. The epidemiological study showed that there was a significantly positive association between maternal hair Ge and NTD risk, with odds ratios (95% confidence interval) of 1.91 (1.24–2.94) and 1.42 (1.00–2.00) for two concerned time-windows covering the early pregnancy period. Likewise, we found a significantly positive dose–response relationship between Ge intake and its concentration in rats’ fur sample. By using a birth cohort, we found that maternal Ge exposure may increase oxidation stress and the likelihood of fever or flu among pregnant women, which were both related to NTD risk. We first concluded that maternal Ge exposure was positively associated with NTD risk with integrated evidence.

Aclonifen could induce implantation failure during early embryonic development through apoptosis of porcine trophectoderm and uterine luminal epithelial cells

Aclonifen is a diphenyl-ether herbicide that is used to control the growth of weeds while growing crops such as corn and wheat. Although the biochemical effects of aclonifen are well characterized, including its ability to inhibit protoporphyrinogen oxidase and carotenoid synthesis, the toxicity of aclonifen in embryonic implantation and development during early pregnancy, has not been reported. Thus, in this study, we investigated the potential interference of aclonifen in embryonic implantation using porcine trophectoderm (pTr) and uterine luminal epithelial (pLE) cells isolated during implantation period of early pregnancy. Cell viability in both pTr and pLE cells significantly decreased in a dose-dependent manner following aclonifen treatment. Moreover, the proportion of cells in the sub-G1 phase of the cell cycle gradually increased upon treatment with increasing concentrations of aclonifen, which in turn led to an increase in the number of apoptotic cells, as determined by annexin V and propidium iodide staining. Aclonifen treatment caused mitochondrial dysfunction by increasing the depolarization of the mitochondrial membrane potential and the mitochondrial calcium concentration. Aclonifen inhibited cell mobility by suppressing the expression of implantation-related genes in pTr and pLE cells. To explore the underlying mechanism, we evaluated the phosphorylation of PI3K and MAPK signaling molecules. The phosphorylation of AKT, S6, JNK, and ERK1/2 were significantly increased by aclonifen. Collectively, our results suggest that aclonifen may interrupt implantation during early pregnancy by disrupting maternal–fetal interaction.

Effects of camel feed supplementation on milk yields and composition during early pregnancy period

A study was conducted between January and March 2022 which was the camel mating season at Ngaremara ward of Isiolo County. The objective was to determine the effect of feed supplementation on milk yield and composition in early pregnancy. It is documented that camels in pastoral environment drop milk production immediately after conception. There is limited information in literature on effects of supplementation on milk production at this stage. A complete randomized design was used to select ten camels which were supplemented with a formulated concentrate supplement made of locally available materials and ten un-supplemented which was the control. All camels were dewormed at the beginning of the study. Formulated feeds were gradually introduced to ten camels by adding 1 kg daily and enticing them with green acacia pods for 2 weeks until all supplemented were able to feed 3.5 kgs in the evening after grazing. Mating took place naturally using a bull between 6th January and 15th February 2022 and one supplemented and two un-supplemented camels were not mated. Milking was done in the mornings after allowing the calves to stimulate the teats for the milk letdown reflex. Production of milk in litres was recorded daily and total production per camel tabulated on weekly basis. Milk samples were taken every week and analyzed for protein and fat levels. Statistical data analysis for milk yields was done by comparing weekly means using ANOVA with the help of SPSS. Mean weekly milk yields per camel in litres for the supplemented and un-supplemented was 8.6067 ± 0.23691 and 8.0797±0.26185 respectively. There was a consistent increase in milk production for the supplemented camels with a significant increase of 1.63 litres at week 3 (P&lt;0.05). The un-supplemented camels had a consistent drop of milk production which was more significant (P&lt;0.05) at week 5 and 7. Supplemented camels milk had percentage increase in fat content of 3.470 ±0.0300 compared to 3.157±0.0228 for the un-supplemented. The supplemented camels were mated earlier than the un-supplemented. Six supplemented and five un-supplemented were confirmed pregnant through the indigenous knowledge method of cocking the tail (kutoa mkia) when the male or a person approaches. The study concludes that feed supplementation can positively affect the lactation curve and fertility in camels. It is recommended that camels be supplemented with feed formulations made from locally available materials at around the time of mating to improve production and reproduction. Further studies are recommended to determine the effect of pregnancy on milk yields and composition.

O8 - Re-defining Antenatal Care - development and implementation of IMAP in a Large Tertiary Referral Maternity Service

Initial Maternity Assessment and Planning – IMAP The staged roll out of a project to offer all women planning to birth at a large tertiary referral maternity service publicly funded first trimester screening had only just begun when the COVID19 pandemic hit in early 2020. What quickly became apparent was the urgent need for change in the way services were delivered and what ensued was the early implementation of First Trimester Screening Service (FTSS) and the development of policy to support the safety of our women, our staff and the wider community. The early implementation of the FTSS saw a number of further developments: 1. Increased rates of reported Domestic violence and or psychosocial vulnerabilities 2. Highlighted duplication of midwifery and obstetric assessments in the FTSS and a traditional booking in visit. 3. Highlighted lack of information available to women regarding models of care and maternity service in general A clinical redesign project, Initial Maternity Assessment and Planning (IMAP) commenced in November 2020 and was implemented in November 2021. The project delivered: 1. A universal entry into the Service via a single electronic referral 2. Detailed patient information regarding the IMAP visit, FTS, pathways and options for models of maternity care provided prior to initial appointment 3. IMAP visit – comprehensive, holistic, multidisciplinary screening assessment of the pregnant woman and baby in the early pregnancy period, detailed risk assessment and risk appropriate management plan and allocated model of care 4. Elements of antenatal care, Low and high risk pathways – schedule of antenatal visits, recommended content and supporting education and resource for women and families. 5. Focussed visits at 16-18 weeks – welcome to your model of care and 36 weeks – Review pregnancy and recommended timing, model and place of birth 6. Comprehensive suite of educational resources accessed via QR code. At this stage the project evaluation is just commencing. We expect to be able to present preliminary outcomes at the conference.